Figure 1.

Monocyte-derived macrophages phagocytose erythrocytes that are opsonized with different anti-RhD IgG1 glycovariants to similar extent. (A) Composition of the bi-antennary glycan at position Asn297 in the IgG Fc domain. Percentages indicated in red are for the unmodified IgG1 variant (14). (B) Erythrocytes opsonized with monoclonal anti-RhD glycovariants (x-axis) were stained with goat-anti-human Ig to determine the amount of antibody deposition. Median fluorescence intensity (MFI) is shown on the y-axis. (C) Phagocytosis of anti-RhD opsonized erythrocytes by monocyte-derived macrophages cultured with GM-CSF (left) or M-CSF (right). Erythrocytes were opsonized with unmodified anti-RhD IgG1 or an anti-RhD IgG1 glycovariant (indicated on the x-axis). Percentage of positive macrophages is shown on the y-axis. (B,C) n = 6–10. Analysis is performed by one-way ANOVA, corrected for multiple comparisons using Dunnett.